Literature summary for 2.7.1.147 extracted from

Castro-Fernandez, V.; Herrera-Morande, A.; Zamora, R.; Merino, F.; Gonzalez-Ordenes, F.; Padilla-Salinas, F.; Pereira, H.M.; Brandao-Neto, J.; Garratt, R.C.; Guixe, V.
Reconstructed ancestral enzymes reveal that negative selection drove the evolution of substrate specificity in ADP-dependent kinases (2017), J. Biol. Chem., 292, 15598-15610 .

Search for more literature for 2.7.1.147

Cloned(Commentary)

Cloned (Comment)	Organism
gene glkA, phylogenetic analysis, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)	Thermococcus litoralis
gene pfkC, phylogenetic analysis, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)	Methanocaldococcus jannaschii
phylogenetic analysis, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)	uncultured bacterium

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified enzyme mutant E72A, hanging drop vapor diffusion method, mixing of 0.002 ml of 8 mg/ml protein in 25 mM HEPES/NaOH, pH 7.8, 30 mM MgCl2, 20 mM fructose 6-phosphate, and 20 mM AMP with 0.002 ml of reservoir solution containing 18% PEG 3350 and 0.15 M NaI, 3 days at 18°C, X-ray diffraction structure determination and analysis at 2.61 A resolution, molecular replacement using the structure of Pyrococcus horikoshii ADP-PFK (PDB ID 3DRW) split into small and large domains as search models	uncultured bacterium

Crystallization (Comment)

Organism

purified enzyme mutant E72A, hanging drop vapor diffusion method, mixing of 0.002 ml of 8 mg/ml protein in 25 mM HEPES/NaOH, pH 7.8, 30 mM MgCl2, 20 mM fructose 6-phosphate, and 20 mM AMP with 0.002 ml of reservoir solution containing 18% PEG 3350 and 0.15 M NaI, 3 days at 18°C, X-ray diffraction structure determination and analysis at 2.61 A resolution, molecular replacement using the structure of Pyrococcus horikoshii ADP-PFK (PDB ID 3DRW) split into small and large domains as search models

uncultured bacterium

Protein Variants

Protein Variants	Comment	Organism
E72A	site-directed mutagenesis, the mutant shows reduced glucokinase and phosphofructokinase activities compared to wild-type enzyme	uncultured bacterium

Inhibitors

Inhibitors	Comment	Organism
D-glucose	-	Methanocaldococcus jannaschii
D-glucose	-	Thermococcus litoralis
additional information	enzyme TlGK is not significantly inhibited by D-fructose 6-phosphate	Thermococcus litoralis

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.0011	-	D-glucose	pH 7.8, 40°C	Methanocaldococcus jannaschii
0.004	-	D-glucose	pH 7.8, 40°C, wild-type enzyme	uncultured bacterium
0.044	-	D-glucose	pH 7.8, 40°C	Thermococcus litoralis
1	-	D-glucose	pH 7.8, 40°C, mutant E72A	uncultured bacterium

Metals/Ions

Metals/Ions	Comment	Organism
Mg2+	required	uncultured bacterium
Mg2+	required	Thermococcus litoralis
Mg2+	required	Methanocaldococcus jannaschii

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.
ADP + D-glucose	uncultured bacterium	-	AMP + D-glucose 6-phosphate	-	?
ADP + D-glucose	Thermococcus litoralis	-	AMP + D-glucose 6-phosphate	-	?
ADP + D-glucose	Methanocaldococcus jannaschii	-	AMP + D-glucose 6-phosphate	-	?
ADP + D-glucose	Methanocaldococcus jannaschii ATCC 43067	-	AMP + D-glucose 6-phosphate	-	?
ADP + D-glucose	Thermococcus litoralis ATCC 51850	-	AMP + D-glucose 6-phosphate	-	?

Organism

Organism	UniProt	Comment	Textmining
Methanocaldococcus jannaschii	Q58999	-	-
Methanocaldococcus jannaschii ATCC 43067	Q58999	-	-
Thermococcus litoralis	Q7M537	-	-
Thermococcus litoralis ATCC 51850	Q7M537	-	-
uncultured bacterium	-	synthetic construct	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, dialysis, and anion exchange chromatography	uncultured bacterium
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, dialysis, and anion exchange chromatography	Thermococcus litoralis
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, dialysis, and anion exchange chromatography	Methanocaldococcus jannaschii

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
ADP + D-glucose	-	uncultured bacterium	AMP + D-glucose 6-phosphate	-	?
ADP + D-glucose	-	Thermococcus litoralis	AMP + D-glucose 6-phosphate	-	?
ADP + D-glucose	-	Methanocaldococcus jannaschii	AMP + D-glucose 6-phosphate	-	?
ADP + D-glucose	-	Methanocaldococcus jannaschii ATCC 43067	AMP + D-glucose 6-phosphate	-	?
ADP + D-glucose	-	Thermococcus litoralis ATCC 51850	AMP + D-glucose 6-phosphate	-	?
additional information	the enzyme also shows phosphofructokinase activity, a bifunctional AncPFK/GK ancestor enzyme	uncultured bacterium	?	-	-
additional information	the enzyme from Methanococcus jannaschii also shows phosphofructokinase activity, a bifunctional MjPFK/GK	Methanocaldococcus jannaschii	?	-	-
additional information	the glucokinase from Thermococcus litoralis shows no activity with fructose 6-phosphate	Thermococcus litoralis	?	-	-
additional information	the enzyme from Methanococcus jannaschii also shows phosphofructokinase activity, a bifunctional MjPFK/GK	Methanocaldococcus jannaschii ATCC 43067	?	-	-
additional information	the glucokinase from Thermococcus litoralis shows no activity with fructose 6-phosphate	Thermococcus litoralis ATCC 51850	?	-	-

Synonyms

Synonyms	Comment	Organism
ADP-dependent glucokinase	-	uncultured bacterium
ADP-dependent glucokinase	-	Thermococcus litoralis
ADP-GK	-	uncultured bacterium
ADP-GK	-	Thermococcus litoralis
ancGK/PFK	-	uncultured bacterium
MjPFK/GK	-	Methanocaldococcus jannaschii
More	see also EC 2.7.1.146	uncultured bacterium
More	see also EC 2.7.1.146	Methanocaldococcus jannaschii
tlGK	-	Thermococcus litoralis

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
40	-	assay at	uncultured bacterium
40	-	assay at	Thermococcus litoralis
40	-	assay at	Methanocaldococcus jannaschii

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
0.24	-	D-glucose	pH 7.8, 40°C, wild-type enzyme	uncultured bacterium
0.3	-	D-glucose	pH 7.8, 40°C, mutant E72A	uncultured bacterium
5	-	D-glucose	pH 7.8, 40°C	Methanocaldococcus jannaschii
42	-	D-glucose	pH 7.8, 40°C	Thermococcus litoralis

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.8	-	assay at	uncultured bacterium
7.8	-	assay at	Thermococcus litoralis
7.8	-	assay at	Methanocaldococcus jannaschii

Cofactor

Cofactor	Comment	Organism
ADP	-	Methanocaldococcus jannaschii
ADP	dependent on	uncultured bacterium
ADP	dependent on	Thermococcus litoralis

Ki Value [mM]

Ki Value [mM]	Ki Value maximum [mM]	Inhibitor	Comment	Organism	Structure
14	-	D-glucose	pH 7.8, 40°C	Thermococcus litoralis
97	-	D-glucose	pH 7.8, 40°C	Methanocaldococcus jannaschii

General Information

General Information	Comment	Organism
evolution	consensus phylogenetic tree of the ADP-dependent sugar kinases family, evolutionary history of enzyme substrate affinity, reconstruction	uncultured bacterium
evolution	consensus phylogenetic tree of the ADP-dependent sugar kinases family, evolutionary history of enzyme substrate affinity, reconstruction	Thermococcus litoralis
evolution	consensus phylogenetic tree of the ADP-dependent sugar kinases family, evolutionary history of enzyme substrate affinity, reconstruction	Methanocaldococcus jannaschii
additional information	molecular modeling, docking with D-glucose and D-fructose 6-phosphate, and molecular dynamics	uncultured bacterium
additional information	molecular modeling, docking with D-glucose and D-fructose 6-phosphate, and molecular dynamics	Thermococcus litoralis
additional information	molecular modeling, docking with D-glucose and D-fructose 6-phosphate, and molecular dynamics	Methanocaldococcus jannaschii

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism
60	-	D-glucose	pH 7.8, 40°C, wild-type enzyme	uncultured bacterium
81	-	D-glucose	pH 7.8, 40°C, mutant E72A	uncultured bacterium
954.54	-	D-glucose	pH 7.8, 40°C	Thermococcus litoralis
4545.45	-	D-glucose	pH 7.8, 40°C	Methanocaldococcus jannaschii